Detection of Lymphoid Markers (CD3 and PAX5) for Immunophenotyping in Dogs and Cats: Comparison of Stained Cytology Slides and Matched Cell Blocks.

Immunolabeling on Romanowsky-stained cytology (RSC) slides can be used, although there is limited evidence of its suitability for phenotyping canine and feline lymphomas. A comparison with matched cell blocks (CB) is missing. Immunolabeling on RSC and CB was compared for lymphoid markers (CD3 and PAX5) in 53 lymphomas and 4 chylous effusions from dogs and cats. The influence of pre-analytical variables (species, time of archive, type of specimens and coverslipping) and the interobserver agreement among the 2 observers was assessed. Fewer CD3+ lymphocytes were identified in RSC, while the PAX5 positivity by RSC and CB had a substantial agreement. Immunodetection of CD3 and the diagnosis of a T-cell population on RSC was more difficult. Lower intensity and higher background were noted in RSC. Immunophenotyping was inconclusive in 54% RSC and 19% CB. The interobserver reproducibility of immunophenotyping on CB was substantial, being higher than in RSC. The immunolabeling performance on the RSC of effusion and feline samples was unsatisfactory. The detection of lymphoid markers, especially membranous antigens in retrospective RSC, is affected by the pre-analytical variables: species, time of the archive, and type of specimens. CB are a more consistent type of sample for immunophenotyping purposes.

Bacterial Isolates from Urinary Tract Infection in Dogs and Cats in Portugal, and Their Antibiotic Susceptibility Pattern: A Retrospective Study of 5 Years (2017-2021).

There are growing concerns regarding the rise of antimicrobial-resistant bacteria in companion animals. This study aimed to bring new insights into the current scenario of Portugal's antimicrobial resistance bacteria isolated from companion animals with urinary tract infections and is the first to be performed during a long period on a large scale. Of a total of 17472 urine samples analyzed, 12,166 (69.6%) (CI 12,200-12,200) were negative for bacterial growth, and 5306 (30.4%) (95% CI 5310-5310) had bacterial growth. Of the culture-positive samples, 5224 (96.6%) (95% CI 5220-5220) were pure cultures and 82 (3.2%) (95% CI 81.9-82.1) had mixed growth. Escherichia coli was the most frequently isolated bacteria (n=2360, 44.5%) (95% CI 2360-2360), followed by Proteus mirabilis (n=585, 11%) (95% CI 583-583), Enterococcus faecium (n=277, 5.2%) (95% CI 277-277) and Staphylococcus pseudintermedius (n=226, 4.3%) (95% CI 226-226). The overall susceptibility rates were low for erythromycin (45.3%) and clindamycin (51.3%), and high for aminoglycosides (96.3%), carbapenems (92.4%), trimethoprim-sulfamethoxazole (81.2%), and quinolones (79.9%). E. coli also showed considerable resistance to amoxicillin-clavulanic acid. The rates of multidrug-resistant bacteria are still high compared to the northern countries of Europe. This study's findings show the emergence of antibiotic resistance in the antibiotic agents commonly used in the treatment of UTIs in dogs and cats in Portugal.

Doing more with less: multiple uses of a single slide in veterinary cytology. A practical approach.

Veterinary cytology faced a remarkable evolution in the last 15 years, in part due to increase recognition of the advantages of the cytology by veterinary clinicians. Simultaneously, there has been a growing awareness by the owners about the importance of a complete diagnostic workup aimed at defining a proper treatment protocol. With the extended use of cytology, challenging diagnostic cases are more frequent, and more clinically useful answers are requested. In this scenario, the use of cytology specimens to perform ancillary techniques is a valid approach. Rather than being simply archived, cytology slides can be a valuable source and a good platform to carry out cytochemistry, immunocytochemistry, and molecular techniques. Therefore, several diagnostic techniques can be applied in tiny samples, thus following the "doing more with less" principle. The aim of this approach is to refine the cytologic diagnosis and provide additional prognostic and therapeutic information. Herein, we detailed this principle in veterinary cytology and reviewed the use of cytology specimens for ancillary techniques as a single procedure, i.e., using the whole slide, or multiple procedures, i.e., multiple procedures applied in the same slide.

Needle rinse cell blocks as an ancillary technique: Diagnostic and clinical utility in gastrointestinal neoplasia.

BACKGROUND: Fine-needle aspirate (FNA) cytology is often the first-choice method for diagnosing gastrointestinal nodular lesions. The FNA material can be converted to histopathology specimens by a needle rinse cell block (NRCB) technique, allowing ancillary studies to refine the cytologic diagnosis. Despite use in human pathology, NRCB has never been applied to canine or feline gastrointestinal neoplasia. OBJECTIVE: This study described NRCB methodology and its diagnostic utility in specific cases of neoplastic gastrointestinal lesions. METHODS: Needle rinses with saline were performed after ultrasound-guided FNAs of two intestinal lymphomas (canine and feline) and a canine gastrointestinal stromal tumor (GIST). The NRCB was prepared using the cell tube block technique and processed for paraffin embedding. Routine immunohistochemistry protocols (using CD3, PAX-5, and Ki-67 for lymphoma cases and vimentin, desmin, S-100, and KIT markers for GIST) were applied to NRCB sections, and the results were compared with matched tissue biopsies. RESULTS: NRCBs with adequate cell numbers, preservation, and good separation of blood were obtained. The diagnosis and immunophenotyping were confirmed in both cases of lymphoma in NRCBs. In the GIST, the immunolabeling of the neoplastic cells in NRCB was completely concordant with the tissue biopsy. CONCLUSIONS: The described methodology is suitable for veterinary settings, having few technical requirements and low invasiveness. The presented cases of gastrointestinal neoplasia highlight the utility of NRCBs as a platform to conduct ancillary studies and refine the cytologic diagnosis.